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Rapid, Ultrasensitive, and Quantitative Detection of SARS-CoV-2 Using Antisense Oligonucleotides Directed Electrochemical Biosensor Chip.
Alafeef, Maha; Dighe, Ketan; Moitra, Parikshit; Pan, Dipanjan.
  • Alafeef M; Bioengineering Department, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
  • Dighe K; Departments of Diagnostic Radiology and Nuclear Medicine and Pediatrics, Center for Blood Oxygen Transport and Hemostasis, University of Maryland Baltimore School of Medicine, Health Sciences Research Facility III, 670 W Baltimore Street, Baltimore, Maryland 21201, United States.
  • Moitra P; Biomedical Engineering Department, Jordan University of Science and Technology, Irbid 22110, Jordan.
  • Pan D; Bioengineering Department, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
ACS Nano ; 14(12): 17028-17045, 2020 Dec 22.
Article in English | MEDLINE | ID: covidwho-880666
ABSTRACT
A large-scale diagnosis of the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) is essential to downregulate its spread within as well as across communities and mitigate the current outbreak of the pandemic novel coronavirus disease 2019 (COVID-19). Herein, we report the development of a rapid (less than 5 min), low-cost, easy-to-implement, and quantitative paper-based electrochemical sensor chip to enable the digital detection of SARS-CoV-2 genetic material. The biosensor uses gold nanoparticles (AuNPs), capped with highly specific antisense oligonucleotides (ssDNA) targeting viral nucleocapsid phosphoprotein (N-gene). The sensing probes are immobilized on a paper-based electrochemical platform to yield a nucleic-acid-testing device with a readout that can be recorded with a simple hand-held reader. The biosensor chip has been tested using samples collected from Vero cells infected with SARS-CoV-2 virus and clinical samples. The sensor provides a significant improvement in output signal only in the presence of its target-SARS-CoV-2 RNA-within less than 5 min of incubation time, with a sensitivity of 231 (copies µL-1)-1 and limit of detection of 6.9 copies/µL without the need for any further amplification. The sensor chip performance has been tested using clinical samples from 22 COVID-19 positive patients and 26 healthy asymptomatic subjects confirmed using the FDA-approved RT-PCR COVID-19 diagnostic kit. The sensor successfully distinguishes the positive COVID-19 samples from the negative ones with almost 100% accuracy, sensitivity, and specificity and exhibits an insignificant change in output signal for the samples lacking a SARS-CoV-2 viral target segment (e.g., SARS-CoV, MERS-CoV, or negative COVID-19 samples collected from healthy subjects). The feasibility of the sensor even during the genomic mutation of the virus is also ensured from the design of the ssDNA-conjugated AuNPs that simultaneously target two separate regions of the same SARS-CoV-2 N-gene.
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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Prognostic study Language: English Journal: ACS Nano Year: 2020 Document Type: Article Affiliation country: Acsnano.0c06392

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Prognostic study Language: English Journal: ACS Nano Year: 2020 Document Type: Article Affiliation country: Acsnano.0c06392